刘玉洋,孙博,赵海誉,丁耀华,张艳,高文雅,田孟尧,王宏洁,周严严,魏晓露,司南,边宝林
LIU Yu-yang,SUN Bo,ZHAO Hai-yu,DING Yao-hua,ZHANG Yan,GAO Wen-ya,TIAN Meng-yao,WANG Hong-jie,ZHOU Yan-yan,WEI Xiao-lu,SI Nan,BIAN Bao-lin

• 1:中国中医科学院中药研究所
• 2:西昌富华金蟾生物科技有限公司

摘要(Abstract)：

为助力解决蟾蜍资源日益减少的窘境，探究蟾蜍中功效成分蟾蜍甾烯类化合物(BDs)生物合成部位及代谢途径，为蟾蜍资源可持续利用提供理论基础。利用UHPLC-Orbitrap-MS对蟾蜍多组织进行检测，筛选BDs在蟾蜍体内的合成部位，并利用质谱成像、匀浆孵化实验加以验证，确定BDs生物合成的关键器官。蟾蜍组织分布结果表明除耳后腺、皮肤组织外，BDs在肝脏中含量最多，胆汁中浓度最高，因此推测肝脏能够生物合成BDs,并通过胆汁代谢分布向蟾蜍全身。DESI-MSI质谱成像检测结果也指出，BDs主要富集在蟾蜍肝脏而并非是未发育成熟的耳后腺。蟾蜍肝脏匀浆孵化证明了蟾蜍肝脏能够独立生物合成BDs。该研究表明中华大蟾蜍在变态、发育、生长过程中，肝脏是蟾蜍体内合成BDs的重要器官，为生物合成BDs以及资源可持续利用提供了有力的理论支撑。
This study explored the biosynthesis of bufadienolides(BDs) in Bufo bufo gargarizans to solve the dilemma of the decreasing resources of B. bufo gargarizans and provide a theoretical basis for the sustainable utilization of the resources. Ultra-high performance liquid chromatography-Orbitrap-mass spectrometry(UHPLC-Orbitrap-MS) was employed to detect the synthesis sites of BDs in B. bufo gargarizans, and the results were verified by desorption electrospray ionization-mass spectrometry imaging(DESI-MSI) and homogenate incubation experiments. BDs in B. bufo gargarizans had the highest content in the liver and the highest concentration in the gallbladder, in addition to the parotid gland and skin, which suggested that the liver could synthesize BDs. The results of DESI-MSI also showed that BDs were mainly enriched in the liver rather than the immature parotid gland. The incubation experiment of liver homogenates demonstrated the liver of B. bufo gargarizans had the ability to synthesize BDs. This study showed that the liver was a major organ for the synthesis of BDs in B. bufo gargarizans during metamorphosis, development, and growth, which provided strong theoretical support for the biosynthesis of BDs and the sustainable utilization of B. bufo gargarizans resources.

关键词(KeyWords)： 蟾蜍;蟾蜍甾烯类成分;组织分布;资源;生物合成
Bufo bufo gargarizans;bufadienolides;tissue distribution;resources;biosynthesis

Abstract:

Keywords:

基金项目(Foundation): 国家林业和草原局重点研发项目(GLM[2021]46号);; 中央级公益性科研院所基本科研业务费专项(ZXKT22004);; 中国中医科学院科技创新工程项目(CL2021A04012)

作者(Author): 刘玉洋,孙博,赵海誉,丁耀华,张艳,高文雅,田孟尧,王宏洁,周严严,魏晓露,司南,边宝林
LIU Yu-yang,SUN Bo,ZHAO Hai-yu,DING Yao-hua,ZHANG Yan,GAO Wen-ya,TIAN Meng-yao,WANG Hong-jie,ZHOU Yan-yan,WEI Xiao-lu,SI Nan,BIAN Bao-lin

DOI: 10.19540/j.cnki.cjcmm.2023105.102

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